CN112753658B - Method for indoor breeding trichogramma dendrolimi by utilizing bombyx camphorata eggs - Google Patents
Method for indoor breeding trichogramma dendrolimi by utilizing bombyx camphorata eggs Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a method for breeding trichogramma dendrolimi trichogramma in a chamber by utilizing bombyx mori eggs, which comprises the steps of killing embryos in the bombyx mori eggs to obtain processed bombyx mori eggs, and breeding the processed bombyx mori eggs by the trichogramma dendrolimus trichogramma: placing the processed camphor silkworm eggs into a container through an opening of the container, and then inoculating the pine moth trichogramma into the container, wherein the pine moth trichogramma is a female bee of the emergence-crossed pine moth; sealing the opening of the container with air permeable material, and culturing in artificial climate box; when the culture time is 24 +/-2 hours, removing the trichogramma dendrolimi from the container; culturing bombyx cinnamomea eggs in a climatic chamber under the culture conditions until the eggs of the trichogramma dendrolimi are eclosized.
Description
Technical Field
The invention relates to an artificial propagation method of insect egg parasitism bees, namely trichogramma dendrolimi trichogramma, in particular to a method for indoor propagation of trichogramma dendrolimi trichogramma by utilizing camphor silkworm eggs, belonging to the technical field of biological control of agricultural pests.
Background
The camphor silkworm (eriogyna (saturnia) pyretoum Westwood) belongs to lepidoptera bombyx bombycidae, is a parasitic pest, is distributed in Zhejiang, Guangdong, Guangxi, Fujian and other places, and uses larva to eat camphor tree leaves. In severe cases, the leaves of the whole tree are eaten, and even the tree dies. The insect is grown in one generation in one year, pupa overwinter in cocoon at branch or bark gap, or forest shrub, etc., emergence begins in next month, and adult can copulate and lay eggs soon after emergence, and has phototaxis. The prevention and control of the camphor silkworms mainly adopt artificial light trapping and killing and chemical agent prevention and control, have less biological prevention and control, mainly use beauveria bassiana powder for prevention and control at the early stage (4 months) of rainy season, at the temperature of more than 24 ℃ and at the relative humidity of more than 95 percent, and have good effect.
Trichogramma (trichogram spp.) has been found for over 100 years to date, can parasitize eggs of various lepidoptera insects, and is the most widely-used egg parasitic natural enemy in biological pest control all over the world. Since 1926, Flanders in the United states successfully propagated trichogramma in large quantities using eggs of the Spodoptera frugiperda cerealella (Olivier), and internationally began mass production of trichogramma hosts using eggs of Spodoptera frugiperda as industrial hosts. In addition, the insecticidal principle of the biological missile product successfully developed by Wuhan virus research institute of Chinese academy of sciences is based on the epidemic of insect viruses, and viruses are transmitted by using egg parasitic wasps (trichogramma) as vectors, so that the initially hatched larvae are diseased, and the purpose of controlling pests by inducing population diseases is achieved.
With the development of sustainable agriculture and the continuous improvement of the requirement of pollution-free green food industry, the biological control technology mainly based on natural enemies is more and more accepted by farmers.
The invention patent with application number 201610145652.3, a method for breeding trichogramma dendrolimi and trichogramma zeae with oak silkworm eggs as hosts, comprises the following steps: under the conditions of 23-29 ℃, 60-90% RH and 12-16L:12-8D of illumination period, simultaneously mixing and inoculating the corn borer trichogramma and the pine moth trichogramma bred by parasitic eggs into the tussah eggs for 24 hours according to the proportion of 4: 1; in the bee breeding process, the ratio of the total number of the inoculated trichogramma to the number of the tussah eggs is controlled to be 5: 1. As the tussah industry is mainly distributed in provinces such as Liaoning, Shandong, Henan, Jilin, Heilongjiang, inner Mongolia, Shanxi and the like in China, the tussah yield in Liaoning province accounts for 80 percent of the national portion and 70 percent of the world portion. In the last 80 th century, corresponding production bases were established in Zhejiang province, but due to the limitation of regions and seasons, the scale production of oak planted in south China and the scale production of squeezed silkworm bred in south China are not achieved. If the northern China oak silkworm egg is adopted for propagation, a refrigeration house needs to be built in Zhejiang province, and the oak silkworm is transported from the northern China to the southern China for storage, so that the mass propagation of the oak silkworm is maintained, and the labor, time and financial investment are high; therefore, the mass production of the local trichogramma breeding in south is difficult to realize. The trichogramma breeding method has the following defects that the parasitic rate and the emergence rate are low, host silkworm eggs are difficult to obtain in summer and autumn, the parasitic types are mostly limited to the trichogramma types in the northeast, and the like.
The mass production process of trichogramma consists mainly of 2 aspects, namely rearing of intermediate hosts and breeding of parasitic wasps, and most production plants concentrate major time and effort for producing intermediate hosts. The selection of the intermediate host suitable for the mass propagation of the trichogramma is a precondition for large-area application of the trichogramma, the mediterranean pink moth eggs and the wheat moth eggs are generally used as the intermediate hosts to carry out large-scale production of the trichogramma abroad, and the mechanized production is realized.
At present, no camphor silkworm egg is used for carrying out mass propagation of trichogramma dendrolimi.
Disclosure of Invention
The invention aims to solve the technical problem of providing a method for breeding a large number of trichogramma dendrolimi trichogramma by utilizing camphor silkworm eggs.
In order to solve the technical problems, the invention provides a method for breeding trichogramma dendrolimi in a chamber by utilizing a bombyx mori egg, which comprises the steps of killing embryos in the bombyx mori egg so as to obtain a processed bombyx mori egg, and breeding the trichogramma dendrolimi trichogramma in the processed bombyx mori egg; the propagation comprises the following steps:
putting the processed camphor silkworm eggs into a container through an opening of the container, and then inoculating trichogramma dendrolimi dendrom limi dendrolimi maxim dendrolimi dendrom limi dendrolimi dendrom limi;
parasitizing the trichogramma dendrolimi on the treated camphor silkworm eggs, and arranging a nutrient source (a supplementary nutrient source) in a container; the nutrient source is used for providing nutrition for the trichogramma dendrolimi;
the number ratio of bee eggs is 2-10: 1;
secondly, sealing the opening of the container by using a breathable material, and then culturing in an artificial climate box; the culture conditions were: the temperature is 25 +/-1 ℃, the relative humidity RH is 75 +/-5%, and the color is completely dark;
when the culture time is 24 +/-2 hours, removing the trichogramma dendrolimi from the container; culturing bombyx cinnamomi eggs in a climatic chamber under the culture condition until the eggs of the trichogramma dendrolimi are eclosized (the culture time is about 10 +/-0.5 days).
Remarks explanation: after the culture in the artificial climate box for 5 days, the inspection shows that the mothball eggs parasitized by the pine moth trichogramma become black, and the eggs not parasitized are still silver gray; after continuously culturing for 10 +/-0.5 days, the adult trichogramma dendrolimi of the new generation starts to emerge continuously (the emergence rate can be as high as 97.7 percent), and can be subjected to subculture.
The improvement of the method for breeding the trichogramma dendrolimi in the chamber by utilizing the eggs of the camphor silkworms of the invention comprises the following steps: directly breeding the trichogramma dendrolimi and trichogramma of the treated camphor silkworm eggs; or firstly storing and then subsequently propagating;
the storage is as follows: placing the processed bombyx cinnamomi egg in a container with air holes, controlling the relative humidity in the container to be 75 +/-5%, and storing the container filled with the bombyx cinnamomi egg at 4 +/-1 ℃.
As a further improvement of the method for indoor propagation of trichogramma dendrolimi by utilizing the bombyx mori eggs, the method for killing embryos in the bombyx mori eggs to obtain the processed bombyx mori eggs comprises the following steps:
placing the camphor silkworm eggs on parchment paper in a natural spawning or artificial egg extruding mode, removing impurities and sterilizing (slight sterilizing), and irradiating for 2 hours under the ultraviolet ray with the light intensity of 21.2Lux to kill embryos in the camphor silkworm eggs to obtain the treated camphor silkworm eggs.
Description of the drawings: the time from spawning/egg extruding to ultraviolet irradiation cannot exceed 24 hours (the air drying after egg extruding is generally controlled to be 3-24 hours). The purpose of the ultraviolet irradiation is to kill the embryos in the eggs of the camphor silkworms.
The invention is further improved by utilizing the method for breeding the trichogramma dendrolimi in the antrodia camphorate egg chamber:
the impurity removal is as follows: blow off (blow off gently) the scale that drops on the camphor-silkworm when artifical crowded ovum on the parchment paper, disinfection (slight disinfection) is: the parchment paper together with the eggs were sterilized by wiping (light wiping) with a cotton ball stained with 75% ethanol.
The invention is further improved by utilizing the method for breeding the trichogramma dendrolimi in the antrodia camphorate egg chamber:
the breathable material is a tampon pledget.
The invention is further improved by utilizing the method for breeding the trichogramma dendrolimi in the antrodia camphorate egg chamber:
the supplementary nutrient source is honey water with the mass concentration of 4-6%; the daily supply of the honey water is as follows: every 100 adult trichogramma dendrolimi bees correspond to 0.2-0.5 ml of honey water.
The method can turn the harmful into the beneficial, and obtains the bombyx mori eggs by the conventional lamp induction, manual removal, manual feeding and other modes, thereby providing excellent hosts in the south as indoor breeding trichogramma.
The invention has the following technical advantages:
1. the invention takes the camphor silkworm eggs as the material, thereby realizing the change of the harm into the benefit and having high eclosion rate; is suitable for breeding trichogramma.
2. The invention provides favorable hosts for the artificial propagation of the southern pine moth trichogramma, the corn borer trichogramma, the borer yellow trichogramma and the like, and lays a foundation for large-scale production.
In conclusion, the invention provides a method for breeding Trichogramma dendrolimi by utilizing the antrodia camphorata egg chamber. The invention takes the eggs of the moths ariogyna (Saturnia) pyretoum Westwood as hosts and can continuously propagate trichogramma dendrolimi for a plurality of generations. The camphor silkworm eggs with embryos killed in the eggs are used as hosts, and in the artificial propagation of trichogramma dendrolimi, the artificial propagation method is not limited by time and space, can be continuously propagated in subculture, has high quality of propagated individuals, stable parasitic rate and high emergence rate, shows various advantages, and provides a new way for biologically preventing and controlling a large number of breeding bees for agricultural and forestry pests.
Drawings
The following describes embodiments of the present invention in further detail with reference to the accompanying drawings.
FIG. 1 is a statistical table of the parasitism rate of trichogramma on tussah eggs (10 d storage) (10 d storage, completely dark climatic chamber);
FIG. 2 is a statistical table of the parasitism rate of trichogramma on eggs of bombyx mori (10 d of preservation) (10 d of preservation, completely dark climatic chamber);
FIG. 3 is a statistical table of the parasitic rate of trichogramma on tussah eggs (fresh) (fresh eggs 0d, completely dark climatic chamber);
FIG. 4 is a statistical table of the parasitic rate of trichogramma on bombyx mori eggs (fresh) (fresh eggs 0d, completely dark climatic chamber);
note: the different lower case letters above the same column of values indicate that P-0.05 level is significantly different.
Detailed Description
The invention will be further described with reference to specific examples, but the scope of protection of the invention is not limited thereto:
example 1, a method for indoor propagation of trichogramma dendrolimi by utilizing bombyx camphorata eggs, sequentially comprising the following steps:
1) collecting and treating camphor silkworms:
the method comprises the steps of artificially collecting bombyx cinnamomea in the first 11 months of the year and inducing collected bombyx cinnamomea moths in the next 2-3 months of the year, and placing the bombyx cinnamomea eggs on parchment paper in an artificial egg squeezing mode for breeding trichogramma.
2) Collecting and treating bombyx mori eggs (treatment of killing embryos in bombyx mori eggs)
Firstly, removing impurities and slightly disinfecting the parchment with the camphor silkworm eggs: slightly blowing off scales on the parchment paper, and slightly wiping the paper surface with a cotton ball stained with 75 percent (volume percent) of ethanol so as to realize slight disinfection treatment; then placing the eggs under the ultraviolet ray with the light intensity of 21.2Lux for 2 hours to kill the embryos in the camphor silkworm eggs; obtaining the processed camphor silkworm eggs.
Description of the drawings: the time from egg extrusion to ultraviolet irradiation cannot exceed 24 hours (preferably 3 to 24 hours).
3) Preservation of Camphor silkworm eggs
Wrapping the parchment paper with the processed bombyx cinnamomi eggs with gauze, placing the parchment paper in a circular plastic box with a small hole drilled in a box cover, placing wet cotton balls in the box for moisturizing (namely ensuring that the humidity in the box is 75-80 percent, so that the cotton balls need to be replaced in time after being dried), and storing the parchment paper in a refrigerator at 4 ℃ for at least 10 days.
4) The propagation of trichogramma dendrolimi comprises the following steps:
taking the preserved camphor silkworm eggs out of the refrigerator, putting the preserved camphor silkworm eggs into a test tube of 1cm multiplied by 5.5cm, and then introducing hatched female bees into the test tube so that the ratio of the eggs to the bees is 2: 1. 9:4, 3:1, 10:3, 4:1, etc. Placing absorbent cotton thread dipped with 5% (mass%) of honey water and sticking the absorbent cotton thread on the inner tube wall of the test tube, and enabling the bottom of the cotton thread to be about 2cm away from the test tube opening to be used as a supplementary nutrient source for adult trichogramma dendrolimi.
The absorbent cotton thread with 5% honey water is replaced every day, so that the daily supply of the honey water is ensured to be: every 100 adult pine moth trichogramma corresponds to 0.2-0.5 ml of honey water.
Secondly, sealing the opening of the test tube by using a degreasing cotton plug, and then culturing in an artificial climate box with the temperature of 25 ℃, the humidity of RH 75 +/-5 percent and complete darkness; when the culture time is 24 hours, removing the female trichogramma dendrolimi from the container; continuously culturing the camphor silkworm eggs in the climatic chamber under the culture condition for 10 days; at the moment, the trichogramma pini trichogrammae eggs are eclosized, namely, the peak formation of the trichogramma pini trichogrammae of the new generation is continuously eclosized, the eclosion rate can reach 97.7 percent, and the secondary feeding can be carried out.
The formula for calculating the emergence rate is as follows: the emergence rate is the number of emerged bees/(number of emerged bees + number of unetched bees) × 100%.
Description of the drawings: when the incubation time is 24h, the female trichogramma dendrolimi is removed from the container, and the parasitism of the bombyx cinnamomea eggs can be observed by sampling. After 5 days of cultivation in the climatic chamber (counting from the time when the female trichogramma dendrolimi wasp is removed from the container), it can be found that: the eggs of the mothball silkworm parasitized by the trichogramma dendrolimi become black, and the eggs which are not parasitized are still silver gray.
Experiment 1:
the method comprises the following steps of (1) taking tussah eggs as hosts, putting egg cards (parchment paper with processed tussah eggs) into a large test tube containing a large amount of female trichogramma of pine moth by a large-bee-quantity rapid inoculation method, wherein the ratio (quantity ratio) of bees to eggs is 10:1 hour later, taking out the egg card, removing the residual bee, and culturing in an incubator at 20 deg.C, 25 deg.C and RH 75 + -5%. Taking ovum from the incubator every 20min for 1 time within 2h, fixing part of ovum in fixing solution for 24h, taking out, and preserving in 80% ethanol; the other part was immediately dissected and viewed in vivo.
After 2h, each fixation and in vivo observation was 1 time per hour;
between 24 and 72 hours, specifically: fixing and observing for 24-50 h, 1 time every 2h, 50-60 h, and 1 time every 0.5 h; fixing and observing for 1 time every 2 hours for 60-72 hours;
after 72h until adult, fix and observe the living body 1 time every 4-6 h.
For convenience, 4 parasitic eggs of bees within different time periods are adopted in the experimental process, so that the embryo body in a certain development period can be repeatedly observed, the experimental work can be carried out in the daytime, and the experimental work cannot be influenced by the rest at night. In the experiment, acetic acid carmine is used for dyeing, and a paraffin section method is adopted for flaking. The results of the experiment are shown in table 1:
TABLE 1 trichogramma in tussah silkworm eggs ontogeny stage
The bombyx cinnamomi egg is used as a host, the egg card (the parchment paper with the processed bombyx cinnamomi egg) is put into a big test tube containing a large amount of female trichogramma of pine moth by a large-bee-quantity inoculation method, the proportion of the bee eggs is also 10:1, other conditions and methods are the same as the above, and the experimental results are shown in table 2:
TABLE 2 trichogramma ontogeny stage in bombyx mori eggs
Note: the trichogramma flies live parasitizing in the parasitic eggs from the eggs until the imagoes eclosion.
From tables 1 and 2, it can be seen that: the trichogramma in 2 silkworm eggs has basically the same ontogeny stage.
Experiment two, parasitic ability comparison of trichogramma to different hosts under indoor conditions:
taking preserved silkworm eggs and tussah eggs of 10 days out of a refrigerator, putting the silkworm eggs into test tubes of 1cm multiplied by 5.5cm, putting about 10 heads of female trichogramma dendrolimi which is inoculated into each test tube, putting the eggs into the test tubes according to different bee-egg ratios (as shown in figures 1 and 2), and adhering absorbent cotton threads which are about 2cm long and are dipped with 5% of honey water to the position, which is about 2cm away from the tube opening, of the inner wall of the test tube to serve as a supplementary nutrient source of adult trichogramma dendrolimi. Sealing with absorbent cotton plug, and culturing in artificial climate box with temperature of 25 deg.C and humidity RH of 75% +/-5% and complete darkness. And after 24 hours, moving out the female trichogramma of the pine moth, and continuously culturing the camphor silkworm eggs and the tussah silkworm eggs in a climatic chamber to wait for observing the parasitic condition.
Experiments are respectively provided with 3-4 repetitions, camphor silkworm eggs and tussah silkworm eggs are sequentially and respectively dissected to observe the parasitic condition of trichogramma, and the egg laying amount of each repetition is counted (an average value is taken).
Description of the drawings: fig. 1 shows the preferred bee-egg ratio parasitism rate when the host is tussah eggs, and fig. 2 shows the preferred bee-egg ratio parasitism rate when the host is camphor silkworm eggs.
The parasitic ratio is calculated by the formula: the parasitic rate is the number of parasitic eggs with eclosion holes/total number of eggs available for parasitization.
The formula for calculating the emergence rate is as follows: the emergence rate is the number of emerged bees/(number of emerged bees + number of unetched bees) × 100%.
The host is bombyx cinnamomi egg, and the eclosion rate is 97.7% at the optimum parasitic ratio (2: 1).
Therefore, it can be seen that: the method can turn harm into benefit, and the parasitic rate of the trichogramma is obviously higher than that of the trichogramma bred indoors by utilizing the excellent host Antrodia camphorata in south as the host, and the emergence rate of the method is also better.
Experiment three, the camphor silkworm egg and the tussah silkworm egg pre-stored for 10 days in experiment two are changed into the camphor silkworm egg and the tussah silkworm egg directly used after treatment (namely, the storage time is 0 day, the storage time is not preserved), and the bee-egg ratio is changed into the one shown in fig. 3 and 4; the rest is equivalent to experiment two.
The results are shown in FIGS. 3 and 4:
by adopting the method, the method comprises the following steps of: under the preferable scheme that the trichogramma pini-chaeta is 2:1, the parasitic rate of the directly used bombyx mori eggs is basically consistent with that of the bombyx mori eggs which are preserved for 10 days.
When the tussah egg is used as a host, the ratio of the tussah egg: under the preferable scheme that the trichogramma pini-chaeta is 1:1, the parasitic rate of the directly used bombyx mori eggs is far greater than that of the bombyx mori eggs which are preserved for 10 days.
Therefore, the effect of the invention is better.
Comparative example 1, the "completely dark artificial climate box" in experiment two (example 1) was changed to "the light cycle was 14L: 10D", and the rest was identical to experiment two;
when the bombyx mori eggs are used as hosts, the mass ratio of the bombyx mori eggs: the parasitic rate of the trichogramma pini-maid is about 95 percent when the trichogramma pini-maid is 2: 1;
when the tussah egg is used as a host, the ratio of the tussah egg: the parasitism rate of trichogramma pini wasp is about 70% when the ratio of trichogramma pini wasp to trichogramma pini wasp is 1: 1.
Finally, it is also noted that the above-mentioned lists merely illustrate a few specific embodiments of the invention. It is obvious that the invention is not limited to the above embodiments, but that many variations are possible. All modifications which can be derived or suggested by a person skilled in the art from the disclosure of the present invention are to be considered within the scope of the invention.
Claims (6)
1. The method for breeding trichogramma dendrolimi in chamber by utilizing the bombyx mori eggs comprises the steps of killing embryos in the bombyx mori eggs so as to obtain the processed bombyx mori eggs, and is characterized in that: breeding the treated camphor silkworm eggs by using the trichogramma dendrolimi trichogramma; the propagation comprises the following steps:
firstly, placing the processed camphor silkworm eggs into a container through an opening of the container, and then inoculating trichogramma dendrolimi to the container, wherein the trichogramma dendrolimi larva female bees which are eclosid and crossed;
parasitizing the trichogramma pini on the treated bombyx mori eggs, and arranging a nutrient source in a container; the nutrient source is used for providing nutrition for the trichogramma dendrolimi;
the number ratio of bee eggs is 2-10: 1;
secondly, sealing the opening of the container by using a breathable material, and then culturing in an artificial climate box; the culture conditions were: the temperature is 25 +/-1 ℃, the relative humidity RH is 75 +/-5%, and the color is completely dark;
when the culture time is 24 +/-2 hours, removing the trichogramma dendrolimi from the container; culturing bombyx cinnamomea eggs in a climatic chamber under the culture conditions until the eggs of the trichogramma dendrolimi are eclosized.
2. The method for indoor reproduction of trichogramma dendrolimi by using bombyx camphorata eggs as claimed in claim 1, which is characterized in that: directly breeding the trichogramma dendrolimi and trichogramma of the treated camphor silkworm eggs; or firstly storing and then subsequently propagating;
the storage is as follows: placing the processed bombyx cinnamomi egg in a container with air holes, controlling the relative humidity in the container to be 75 +/-5%, and storing the container filled with the bombyx cinnamomi egg at 4 +/-1 ℃.
3. The method for indoor propagation of trichogramma dendrolimi by utilizing bombyx mori eggs according to claim 1 or 2, wherein the method for killing embryos in bombyx mori eggs to obtain the processed bombyx mori eggs comprises the following steps:
placing the camphor silkworm eggs on parchment paper by adopting a natural egg laying or artificial egg squeezing mode, removing impurities and sterilizing the parchment paper with the camphor silkworm eggs, and irradiating for 2 hours under the ultraviolet ray with the light intensity of 21.2Lux to kill embryos in the camphor silkworm eggs to obtain the treated camphor silkworm eggs.
4. The method for indoor reproduction of trichogramma dendrolimi by utilizing eggs of bombyx mori as claimed in claim 3, which is characterized in that:
the impurity removal is as follows: blow away the scale that drops on the camphor-silkworm when artifical crowded ovum on the parchment paper, the disinfection is: the parchment paper and eggs are disinfected by wiping with a cotton ball stained with 75% ethanol.
5. The method for indoor reproduction of trichogramma dendrolimi by utilizing bombyx mori eggs as claimed in claim 4, wherein:
the breathable material is a tampon pledget.
6. The method for indoor reproduction of trichogramma dendrolimi by utilizing bombyx mori eggs as claimed in claim 4, wherein:
the nutrient source is a supplementary nutrient source, and the supplementary nutrient source is honey water with the mass concentration of 4-6%; the daily supply of the honey water is as follows: every 100 adult pine moth trichogramma corresponds to 0.2-0.5 ml of honey water.
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| CN102823550A (en) * | 2012-09-19 | 2012-12-19 | 吉林农业大学 | Light environment control method for breeding trichogramma dendrolimi with oak silkworm eggs |
| CN105766803A (en) * | 2016-03-15 | 2016-07-20 | 吉林农业大学 | Method for blended breeding of trichogramma dendrolimi and trichogramma ostriniae with oak silkworm eggs as hosts |
| CN110742022A (en) * | 2019-10-24 | 2020-02-04 | 山东省农业科学院植物保护研究所 | Trichogramma collection device that lays eggs |
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